ABSTRACT HIV/AIDS continues to be a major global health problem, with a worldwide estimate by the CDC of 38 million people with HIV. HIV viral load (VL) test measures the number of HIV-1 RNA per mL of blood. The test is a marker of antiretroviral treatment (ART) efficacy and adherence to treatment, which is critical for disease management. About 22 million people with HIV are on ART; however, existing HIV VL tests are not suitable for use at the point-of-care (POC) in resource limited settings. These systems are expensive, require reliable electricity, and skilled operator training. Hence, there is an urgent demand for HIV VL testing at POC to decentralize ART monitoring. The goal of this SBIR Phase I project is to demonstrate feasibility of rapid HIV-1 VL quantification from a finger stick, using methods suitable for POC. This project combines Redbud Labs's microfluidic solutions enabling faster, smaller molecular testing with the HIV detection expertise of the Diagnostics Program at PATH (Seattle, WA). PATH has demonstrated a reverse transcriptase Recombinase Polymerase Amplification (RT-RPA) assay to rapidly amplify HIV-1 RNA and DNA. This assay is dependent on efficient mixing for maximum sensitivity and accuracy. We will combine PATH's work on HIV-1 detection with our microfluidic mixer (MXR) and novel blood tool for blood fractionation. We will demonstrate each step of the protocol from sample processing (i.e., blood stabilization, fractionation, and nucleic acid extraction) to detection on-cartridge. The successful completion of this project will position us to develop a sample-to-answer system in Phase II. The public health impact of this work is significant in the effort to meet the UNAIDS goal of 90% viral suppression by 2020.